Van Wonterghem Elien, Van Hoecke Lien, Van Imschoot Griet, Verhaege Daan, Burgelman Marlies, Vandenbroucke Roosmarijn E
VIB Center for Inflammation Research, VIB; Department of Biomedical Molecular Biology, Ghent University.
VIB Center for Inflammation Research, VIB; Department of Biomedical Molecular Biology, Ghent University;
J Vis Exp. 2022 Dec 16(190). doi: 10.3791/64733.
The choroid plexus (CP), a highly vascularized structure protruding into the ventricles of the brain, is one of the most understudied tissues in neuroscience. As it is becoming increasingly clear that this tiny structure plays a crucial role in health and disease of the central nervous system (CNS), it is of utmost importance to properly dissect the CP out of the brain ventricles in a way that allows downstream processing, ranging from functional to structural analysis. Here, isolation of the lateral and fourth brain ventricle mouse CP without the need for specialized tools or equipment is described. This isolation technique preserves the viability, function, and structure of cells within the CP. On account of its high vascularization, the CP can be visualized floating inside the ventricular cavities of the brain using a binocular microscope. However, transcardial perfusion required for downstream analysis can complicate the identification of the CP tissue. Depending on the further processing steps (e.g., RNA and protein analysis), this can be solved by visualizing the CP via transcardial perfusion with bromophenol blue. After isolation, the CP can be processed using several techniques, including RNA, protein, or single cell analysis, to gain further understanding on the function of this special brain structure. Here, scanning electron microscopy (SEM) on whole mount CP is used to get an overall view of the structure.
脉络丛（CP）是一种高度血管化的结构，突出于脑室中，是神经科学领域研究最少的组织之一。随着越来越清楚地认识到这个微小结构在中枢神经系统（CNS）的健康和疾病中起着至关重要的作用，以一种允许进行从功能分析到结构分析等下游处理的方式，从脑室中正确分离出脉络丛至关重要。在此，描述了一种无需专门工具或设备即可分离小鼠侧脑室和第四脑室脉络丛的方法。这种分离技术保留了脉络丛内细胞的活力、功能和结构。由于其高度血管化，使用双目显微镜可以看到脉络丛漂浮在脑室腔内。然而，下游分析所需的经心腔灌注会使脉络丛组织的识别变得复杂。根据进一步的处理步骤（如RNA和蛋白质分析），这可以通过用溴酚蓝进行经心腔灌注来可视化脉络丛来解决。分离后，脉络丛可以使用多种技术进行处理，包括RNA、蛋白质或单细胞分析，以进一步了解这个特殊脑结构的功能。在此，对整个脉络丛进行扫描电子显微镜（SEM）观察以获得该结构的整体视图。
